The PhosphoPlus® MKK3/MKK6 (Ser189/207) Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of MKK3/MKK6 at Ser189/207.
Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9) Rabbit mAb recognizes endogenous levels of MKK3 and MKK6 proteins only when phosphorylated at Ser189 (MKK3) or Ser207 (MKK6). MKK3 (D4C3) Rabbit mAb recognizes endogenous levels of total MKK3 protein. This antibody cross-reacts slightly with MKK6 but demonstrates an approximate 10-fold preference for MKK3 using recombinant, purified proteins.
Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser189 of human MKK3 protein and Ser207 of human MKK6 protein and with a synthetic peptide corresponding to residues near the carboxy terminus of human MKK3 protein.
MKK3 and MKK6 are two closely related dual-specificity protein kinases that activate p38 MAP kinase (1-5). MKK3 and MKK6 both phosphorylate and activate p38 MAP kinase at its activation site, Thr-Gly-Tyr, but do not phosphorylate or activate Erk1/2 or SAPK/JNK. Phosphorylation of p38 MAP kinase dramatically stimulates its ability to phosphorylate protein substrates such as ATF-2 and Elk-1. MKK3 and MKK6 are both activated by different forms of cellular stress and inflammatory cytokines (4,5). Activation of MKK3 and MKK6 occurs through phosphorylation at Ser189 and Thr222 on MKK3 (2) and Ser207 and Thr211 on MKK6 (4,5).
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